Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17th November 1997 - 28th November 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Test material form:
solid: bulk
Details on test material:
Buff-colored
Specific details on test material used for the study:
Sponsor's identification: UK-114,958
Batch number: 116044/D/16/X2/1
Date received: 28 October 1997
Description: buff solid
Storage conditions: room temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Three New Zealand White rabbits, supplied by David Percival Ltd, Moston, Sandbach, Cheshire, UK, were used. At the start of the study the animals weighed 2.59 to 2.81 kg and were twelve to sixteen weeks old. After a minimum acclimatisation period of five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended metal cages. Free access to mains drinking water and food (STANRAB SQC Rabbit Diet, Special Diets Services Ltd, Witham, Essex, UK) was allowed throughout the study.

The animal room was maintained at a temperature of 16 to 20°C and relative humidity of 50 to 65%. On one occasion the temperature was below the limit specified in the protocol (17°C). This deviation was considered not to affect the purpose or integrity of the study. The rate of air exchange was approximately fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light and twelve hours darkness.

Test system

Controls:
yes
Amount / concentration applied:
One rabbit was initially treated. A volume of 0.1 ml of the test material, which was found to weigh approximately 66 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test material, an assessment of the initial pain reaction was made.
Duration of treatment / exposure:
Single exposure, eyelids held together for about second immediately after application.
Observation period (in vivo):
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

An additional observation was made in one treated eye on Day 7 to assess the reversibility of the ocular effects.
Number of animals or in vitro replicates:
3 rabbits
Details on study design:
Three New Zealand White rabbits, supplied by David Percival Ltd, Moston, Sandbach, Cheshire, UK, were used. At the start of the study the animals weighed 2.59 to 2.81 kg and were twelve to sixteen weeks old. After a minimum acclimatisation period of five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended metal cages. Free access to mains drinking water and food was allowed throughout the study.

The animal room was maintained at a temperature of 16 to 20°C and relative humidity of 50 to 65%. On one occasion the temperature was below the limit specified in the protocol (17°C). This deviation was considered not to affect the purpose or integrity of the study. The rate of air exchange was approximately fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light and twelve hours darkness.

Procdure:
Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Animals showing evidence of ocular lesions were rejected and replaced.

One rabbit was initially treated. A volume of 0.1 ml of the test material, which was found to weigh approximately 66 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test material, an assessment of the initial pain reaction was made.

After consideration of the ocular responses produced in the first treated animal, two additional animals were treated. In order to minimise pain on application of the test material, one drop of local anaesthetic ("Ophthaine", 0.5% proxymetacaine hydrochloride, E R Squibb & Sons Limited, Hounslow, Middlesex, UK) was instilled into both eyes of these animals 1 to 2 minutes before treatment.

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation given in Appendix I, (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).

Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

An additional observation was made in one treated eye on Day 7 to assess the reversibility of the ocular effects.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24 h
Score:
ca. 10
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
48 h
Score:
ca. 5
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
72 h
Score:
ca. 5
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
7 d
Score:
ca. 0
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24 h
Score:
ca. 40
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
48 h
Score:
ca. 80
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24 h
Score:
ca. 40
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
48 h
Score:
ca. 80
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24 h
Score:
ca. 5
Irritation parameter:
iris score
Basis:
animal #1
Time point:
48 h
Score:
ca. 0
Irritation parameter:
iris score
Basis:
animal #1
Time point:
72 h
Score:
ca. 0
Irritation parameter:
iris score
Basis:
animal #1
Time point:
7 d
Score:
ca. 0
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24 h
Score:
ca. 5
Irritation parameter:
iris score
Basis:
animal #2
Time point:
48 h
Score:
ca. 5
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24 h
Score:
ca. 5
Irritation parameter:
iris score
Basis:
animal #3
Time point:
48 h
Score:
ca. 5
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24 h
Score:
ca. 10
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
48 h
Score:
ca. 8
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
72 h
Score:
ca. 4
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
7 d
Score:
ca. 0
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24 h
Score:
ca. 18
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
48 h
Score:
ca. 18
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24 h
Score:
ca. 16
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
48 h
Score:
ca. 20
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
24 h
Score:
ca. 25
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
48 h
Score:
ca. 13
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
72 h
Score:
ca. 9
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
7 d
Score:
ca. 0
Irritation parameter:
overall irritation score
Basis:
animal #2
Time point:
24 h
Score:
ca. 63
Irritation parameter:
overall irritation score
Basis:
animal #2
Time point:
48 h
Score:
ca. 103
Irritation parameter:
overall irritation score
Basis:
animal #3
Time point:
24 h
Score:
ca. 61
Irritation parameter:
overall irritation score
Basis:
animal #3
Time point:
48 h
Score:
105
Other effects:
A dulling of the normal lustre of the cornea was noted in one treated eye at the 1-hour observation. Diffuse corneal opacity was noted in this treated eye at the 24, 48 and 72-hour observations. Iridial inflammation was noted in this treated eye at the 1 and 24-hour observations. Moderate conjunctival irritation was noted in this treated eye at the 1, 24 and 48-hour observations with minimal conjunctival irritation at the 72-hour observation. This treated eye appeared normal at the 7-day observation.

Any other information on results incl. tables

Residual test material was noted around the treated eye of all animals at the 1 -hour observation.

A dulling of the normal lustre of the cornea was noted in two treated eyes at the 1-hour observation. Areas of translucent corneal opacity were noted in these two treated eyes at the 24-hour observation. Opaque opacity over approximately one quarter of the lower cornea with translucent or opalescent opacity over the remaining area was noted in these two treated eyes at the 48-hour observation. Iridial inflammation was noted in these two treated eyes at the 1, 24 and 48-hour observations. Moderate conjunctival irritation was noted in these two treated eyes at the 1-hour observation with severe conjunctival irritation at the 24 and 48-hour observations.

These two animals showed signs of discomfort and were therefore killed for humane reasons immediately after the 48 -hour observation.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
The test material, UK-114,958, produced a maximum group mean score of 73.7 and was classified as AT LEAST A SEVERE IRRITANT (CLASS 6 ON A 1 TO 8 SCALE) to the rabbit eye according to a modified Kay and Calandra classification system. The test material was also considered to be irritant according to EU labelling regulations.
Executive summary:

A study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion" (adopted 24 February 1987) and Method B5 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 93/21/EEC) relating to the classification, packaging and labelling of dangerous substances.

A single application of the test material to the non-irrigated eye of three rabbits produced diffuse or opaque corneal opacity, iridial inflammation and moderate to severe conjunctival irritation. Dulling of the normal lustre of the cornea was noted in all treated eyes at the 1-hour observation. Two animals showed signs of discomfort and were therefore killed for humane reasons immediately after the 48- hour observation. One treated eye appeared normal at the 7-day observation.

The test material produced a maximum group mean score of 73.7 and was classified as at least a severe irritant (Class 6 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. The test material was also considered to be irritant according to EU labelling regulations.